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Journal of Veterinary Science & Animal
Husbandry
Volume 1 | Issue 3
Research Article

Open Access

Anti-Inflammatory and Anti-Arthritic Efficacy and Safety of Purified
Shilajit in Moderately Arthritic Dogs
Lawley S1, Gupta RC*1, Goad JT1, Canerdy TD1 and Kalidindi SR2
Murray State University, Breathitt Veterinary Center, Hopkinsville, KY,
USA
2
Natreon Inc. New Brunswick, NJ,
USA
1

*Corresponding author: Gupta RC, Professor and Head of Toxicology Department, Murray State
Univer- sity, Breathitt Veterinary Center, P.O. Box 2000; 715 North Drive, Hopkinsville, KY, 42240-2000,
USA; Fax: (270) 886-4295, Tel: (270) 886-3959, E-mail: rgupta@murraystate.edu
Citation: Lawley S et al (2013) Anti-Inflammatory and Anti-Arthritic Efficacy and Safety of Purified
Shilajit in Moderately Arthritic Dogs. J Vet Sci Anim Husb 1: 302
Received Date: October 31, 2013 Accepted Date: December 12, 2013 Published Date: December 16,
2013

Abstract
The objective of this investigation was to evaluate the efficacy and safety of purified Shilajit in moderately arthritic dogs. Ten
client- owned dogs in a randomized double-blinded study received either a placebo or Shilajit (500 mg) twice daily for a period of
five months. Dogs were evaluated each month for physical condition (body weight, body temperature, heart rate, and respiration
rate) and pain associated with arthritis (overall pain, pain from limb manipulation, and pain after physical exertion). Serum
samples collected from these dogs were examined each month for biomarkers of liver (bilirubin, ALT, and AST), kidney (BUN and
creatinine) heart and mus- cle (creatine kinase) functions. The findings of this study revealed that dogs receiving Shilajit (Group-II)
showed a significant (P<0.05) reduction in pain from limb manipulation by day 60, and overall pain and pain after physical
exertion by day 120. Maximum pain reduction, using all three criteria, was observed on day 150. Pain level remained significantly
unchanged in dogs receiving the placebo. Dogs in either group showed no significant change (P>0.05) in physical parameters or
serum markers, suggesting that Shilajit was well tolerated by moderately arthritic dogs. It was concluded that Shilajit significantly
(P<0.05) reduced pain in osteoarthritic dogs and markedly improved their daily life without any side effects.

Keywords:

Purified Shilajit; Osteoarthritis

in canine; Shilajit safety; Anti-arthritic

nutraceutical

Introduction
Currently, there are more than 78 million dogs in the US,
and every fifth adult dog suffers from arthritis [1]. Dogs
suffer more frequently from osteoarthritis (a chronic and
progres- sive disease) than any other form of arthritis.
Osteoarthritis, also known as degenerative joint disease, is a
functional disor- der of the joint, characterized by a change
in joint shape sec- ondary to a loss of articular cartilage,
osteophyte formation, subchondral sclerosis, bone marrow
lesions and synovial pro- liferation, with subsequent
alteration of mechanical proper- ties that result in
decreased stability, movement and loading [2-9]. In the early
stages of osteoarthritis, there is a progressive depletion of
the cartilage proteoglycan leading to a net loss of matrix
from the cartilage [10]. This leads to a cascade of negative
events, including changes in enzymatic cleavage of
proteoglycans and an increase in minor collagen types
www.annexpublishers.com

leading to structural damage and deterioration of the
cartilage. The breakdown of cartilage can be increased with
certain enzymes including the matrix metalloproteinase
enzymes (MMPs). The severity of cartilage lesions in
osteoarthritis can be cor-

Volume 1 | Issue
3
Journal of Veterinary Science & Animal
related with the levels of collegenase (MMP-1) present [7Husbandry
9]. The cascading events lead to increased friction and
inflamma- tion in the joints. Dogs suffering with
osteoarthritis show stiff- ness of joints, crepitus, pain upon
manipulation of the joint, periarticular swelling, palpable
effusion, restriction in range of motion, lameness and pain
and loss of function [7, 11-16]. The combination of history,
physical exam, and radiograph can help diagnose
osteoarthritis in a canine patient. Radiographs indicate
osteophytosis and subchondral bone sclerosis that de- velops
over time in osteoarthritic patients. MRI findings can reveal
changes consistent with osteoarthritis in the cartilage
[17,18].

2

There are multiple factors which influence dogs to develop
ar- thritis, such as genetic predisposition, trauma or injury to
the bones or joints, aging, poor nutrition, obesity and
environmen- tal factors [1,19]. Often, large breed dogs (e.g.
German Shep- herds, Labrador Retrievers, Siberian Huskies,
Rottweilers and others) are more prone to develop
osteoarthritis than are small breeds [20-22].

www.annexpublishers.com

Volume 1 | Issue
3
Arthritic dogs show a variety of signs and symptoms, from
limping to an inability to walk due to joint stiffness. Pain
is the number one complaint [11-14]. The objectives in
treating arthritis are to minimize joint pain by reducing the
inflamma- tion and slowing the progression of the cartilage
damage and to increase joint flexibility. These goals are
achieved by employing a variety of pharmaceuticals,
nutraceuticals, disease modify- ing agents and physical
therapy along with acupuncture [14,
23-26]. Cyclooxygenase (COX) inhibiting non-steroidal antiinflammatory drugs (NSAIDS, such as carprofen,
deracoxib, etodolac, firocoxib, ketoprofen, meloxicam,
robenacoxib, rofecoxib, tepoxalin, etc) are used with caution
as they may cause some adverse effects, such as reduced
appetite, vomiting, gastrointestinal bleeding and hepatic and
renal damage and dysfunction [23, 27-32]. In a recent
prospective (randomized, placebo-controlled, blinded) study,
Monteiro-Steagall et al [33] found that the incidence of
adverse effects was not statistically different between treated
and control dogs.
Recently, dog owners and veterinarians have considered
man- aging osteoarthritis in dogs with herbs and
nutraceuticals. Today, in humans and animals, glucosamine
and chondroitin sulfate are the most common nutraceuticals
to provide build- ing blocks to repair the cartilage and to
minimize and slow down the progression of osteoarthritis to
ease inflammatory pain [24,25, 34-37].
Shilajit is a blackish brown exudate from the sedimentary
rocks of the pristine himalayan mountains at about 10,000
ft altitude. Purified shilajit is prepared from this exudate by a
proprietary extraction process. the main constituents of
shila- jit are
dibenzo-α-pyrones(DBPs),
DBPchromoproteins and fulvic acids with DBP core. On the
other hand, alluvial(plant based) fulvic acids do not have
DBP core and do not have the same therapeutic efficacy as
shjilajit. In fact, alluvial fulvic ac- ids are used as fertilizers.
Shilajit exerts many important roles in biological systems:
(1) improves bioavailability of minerals and nutrients, (2)
provides electrolytes, (3) detoxifies toxic sub- stances,
including heavy metals, (4) improves immune system and (5)
exhibits antioxidant properties. Shilajit has been used for
more than 3000 years for both preventative health and for
treating many diseases in humans, including diabetes,
arthritis, hypertension, immune dysfunction and loss of
memory [3840]. The present study was undertaken to discover the therapeutic efficacy and
safety of purified Shilajit in
moderately arthritic dogs. The hypothesis of the present
investigation was that purified Shilajit will provide aniinflammatory and anti- arthritic effects in moderately
arthritic dogs with minimal side effects. In this paper, we
present the findings of Shilajit as an ef- fective and safe
supplement that exerts anti-inflammatory and anti-arthritic
effects in osteoarhtritic dogs. Shilajit appears very promising
and offers an alternative to many other previously reported
pharmaceuticals and nutraceuticals.

Materials and methods
Animals
40-65 pounds, were used in this study. These dogs, based on
signs of joint stiffness, lameness and radiographic evidence,
had pain at the level of moderate arthritis. Our inclusion
cri- teria of dogs for this study excluded those having liver,
kidney or heart disease, neoplasia, cancer or any other major
disease. Institutional Animal Care and Use Committee
(IACUC) ap- proval and owner consents were obtained prior
to the initia- tion of this study.

Experimental design
In a randomized double-blinded study, ten client-owned dogs
received a placebo (Group-I) or Shilajit (Group-II). The placebo group contained only two dogs because IACUC
advised us to keep to the number of dogs to a minimum due
to the fact that arthritic dogs suffer from pain for a period of
five months. Moreover, findings from our previous studies
revealed that dogs receiving placebo showed no
improvement in arthritis pain [11-14]. Eight dogs in GroupII received purified Shilajit (500 mg) twice daily (one
capsule before morning meal and one capsule before
evening meal) for a period of five months. None of the dogs
received any treatment or supplement for 3 to
4 weeks prior to the study or during the study period.

Pain measurement
At pre-determined intervals (i.e. 30 days), each dog was
Ten client-owned moderately arthritic dogs, weighing between

evalu- ated for overall pain, pain upon limb manipulation and
pain af- ter physical exertion, for a period of five months.
Overall pain, on a scale of 1-10, was graded as: 0, no pain:
2.5, mild pain: 5, moderate pain: 7.5, severe pain: 10, severe
and constant pain. Pain after limb manipulation, on a scale
of 0-4, was evaluated as: no pain: 0, mild pain: 1, moderate
pain: 2, severe pain: 3, severe and constant pain: 4. Pain after
physical exertion, on a scale of 0-4, was evaluated as: no
pain: 0, mild pain: 1, moder- ate pain: 2, severe pain: 3, and
severe and constant pain: 4. The physical examination of
each limb started with the forelimbs and ended with the rear
limbs. The evaluation focused on ma- nipulation of the limbs
in a forward, backward and circular motion. Three main
joints in each limb included for evalua- tion were the
shoulder joint, knee joint and stifle joint. Pop- ping and
cracking of the joint as well as vocal pain were noted for each
canine. Detailed criteria of the measurement of pain are
provided in our earlier publications [11-14]. The present
investigation was carried out on moderately arthritic dogs. A
moderately arthritic dog exhibits overall pain of about 5 on a
scale of 1-10; pain upon limb manipulation about 2 on a
scale of 1-4; and pain after physical exertion about 2 on a scale
of 1-4.

Physical examination
On a monthly basis, dogs were given a full physical examination for body weight, body temperature and heart rate.
Normal values for these parameters are mentioned below
Table 1.
3

Journal of Veterinary Science & Animal
Husbandry

Parameter

Unit

Placebo/Treated

Day 0

Day 30

Day 60

Day 90

Day 120

Day 150

Body Weight

lb

Placebo

54.1±11.05

54.0±10.8

53.4±9.85

54.6±8.95

53.6±9.95

53.7±9.5

Treated

54.5±3.37

54.9±3.53

54.4±3.26

54.9±3.16

54.5±3.22

54.9±3.27

Heart Rate

Beats/min

Placebo

135±15

120±0

115±5

90±10

98±2

99±1

Treated

106.8±4.96

117.6±5.89

96.5±6.47

102.4±3.40

102.8±4.60

108.9±2.81

Body Temperature

ºF

Placebo

100.3±5.41

100.3±0.2

99.9±0.65

100.3±0.2

100.4±0.3

100.1±1.2

Treated

100.1±0.43

99.6±0.26

99.6±0.23

99.6±0.29

100.9±0.30

100.4±0.20

Values are means±SEM; Normal heart rate= 70-160 beats per minute; Normal body temperature= 101-102.5 ºF.

Table 1: Effects of placebo or Shilajit (500 mg, bid) on physical parameters in moderately arthritic dogs

Parameter

Unit

Placebo/Treated

Day 0

Day 30

Day 60

Day 90

Day 120

Day 150

BUN

mg/dl

Placebo

19.5±3.5

18±1

22±2

21±1

17±1

17±4

Treated

17.1±2.88

15.6±1.78

18.1±2.30

18.1±2.30

20.5±2.85

20.8±2.64

Placebo

0.92±0.16

0.75±0.02

0.72±0.02

0.76±0.09

0.81±0.02

0.82±0.03

Treated

1.00±0.09

0.79±0.03

0.79±0.04

0.79±0.09

0.92±0.05

0.94±0.04

Placebo

0.5±0.3

0.2±0

0.3±0.1

0.35±0.05

0.2±0

0.3±0.1

Treated

0.21±0.04

0.25±0.04

0.23±0.05

0.39±0.09

0.26±0.04

0.31±0.04

Placebo

45.5±23.5

35.5±8.5

28±0

40±12

27±0

57.5±14.5

Treated

47.9±4.54

59.9±8.41

75.1±20.4

57.6±4.59

60.8±7.36

58.6±4.95

Placebo

127.5±39.5

123±33

84±4

104±15

779.5±629.5

166.4±25.7

Treated

198.4±44.7

349±80.9

224.6±66.2

132.0±28.2

246.0±84.8

166.4±25.7

Placebo

28.5±3.5

22±7

19.5±5.5

23.5±5.5

45.5±26.5

92.5±3.5

Treated

27.1±3.26

31.5±3.13

28.4±3.57

28.5±2.35

30.5±3.59

29.1±3.60

Creatinine

mg/dl

Total bilirubin mg/dl
ALT

IU/L

AST

IU/L

CK

IU/L

Values are means±SEM; Normal reference values: BUN (7-26 mg/dl); Creatinine (0.0-1.35 mg/dl); Total bilirubin (0.1-0.6 mg/dl); ALT (10-120 IU/L); AST (15-65 IU/L); CK (60-450
IU/L).

Table 2: Effects of placebo or Shilajit (500 mg, bid) on serum biomarkers of liver (bilirubin, ALT, and AST), kidney (BUN and creatinine), heart and
skeletal muscle (CK) functions in moderately arthritic dogs

Serum biomarkers assays
Blood samples were collected from the cephalic vein using
a 3 mL syringe with a 22 gauge needle and were stored in a
marble top tube. Samples were then spun to collect serum
and transferred to a red top tube for evaluation. Serum
samples were collected each month and analyzed for liver
(bilirubin, ALT, AST), kidney (BUN and creatinine), heart
and muscle (CK) functions, using a Beckman AU 480 serum
analyzer. The serum sample assay indicated that neither
placebo nor Shila- jit produced adverse effects in vital
organs of arthritic dogs. Normal reference values of all
serum parameters are provided below Table 2.

Statistical analysis
The data presented are means ± SEM. Statistical significance
of difference was determined by analysis of variance
(ANOVA) coupled with Tukey-Kramer test (P<0.05) using
the NCSS2000
Statistical Analysis and Graphics Software for Windows®.

Results
In this double-blinded clinical trial, moderately arthritic
dogs receiving placebo (Group-I) or purified Shilajit (500 mg)

twice
Journal of Veterinary Science & Animal
daily for a period of five months, were evaluated for
Husbandry
physical parameters, pain level and serum biomarkers of
liver, kidney, heart and muscle functions.
On a monthly basis, each dog was examined for pain level
(overall pain, pain after limb manipulation and pain
after physical exertion) and the data are shown in Figures 1-3.
While evaluating overall pain, the key points were to observe
the dog’s gait, range of motion, ability to sit or lie down,
ability to rise from a sitting position, ability to rise from a
lying position, and posture while standing. Group-II dogs
receiving Shilajit showed significant (P<0.05) reduction in
overall pain by day
120 (3.00±0.27) compared to day 0 (5.38±0.75). The
maximum reduction in overall pain was noted on day 150
(2.38±0.32). Dogs receiving placebo showed no significant
change in overall pain.
Pain after limb manipulation was measured in each limb
of the dog (placed in lateral recumbency) for flexibility,
joint integrity, crepitance and vocalization. The pain level
was sig- nificantly reduced by day 60 (1.63±0.18) and the
trend contin- ued until day 150 when the pain level was
maximally reduced (1.25±0.31) (Figure 2). Pain after limb
manipulation remained significantly unchanged in Group-1
(placebo) dogs. Figure 3 shows the data of pain after physical
exertion in dogs receiving placebo or Shilajit. The canines
were evaluated for pain after three minutes of jogging. After
jogging, pain level was assessed

4
based on the dog’s unwillingness to move, body position,
limp- ing, flexibilty, joint integrity, crepitance and
vocalization. Sig- nificant reduction in pain after physical
exertion was noted on day 120 (1.37±0.26) compared to day
0 (2.63±0.26) in dogs re- ceving Shilajit. The maximum
reduction in pain was observed on day 150 (1.25±0.28).
Dogs on placebo showed no signifi- cant change in pain
level.

*Significantly
(P<0.05).

different

compared

to

pre-treated

values

Figure 1: Effects of placebo or Shilajit (500 mg, bid) on overall pain in
mod- erately arthritic dogs. Overall pain was graded on a scale of 0-10 (0,
no pain;
2.5, mild pain; 5, moderate pain; 7.5, severe pain; and 10, severe and
constant pain). Details of overall pain measurement criteria are described
in the text and our previous publications [11-14].

*Significantly
(P<0.05).

different

compared

to

pretreated

values

Figure 2: Effects of placebo or Shilajit (500 mg, bid) on pain from limb
manip- ulation in moderately arthritic dogs. Pain from limb manipulation
was graded on a scale of 0-4 (0, no pain; 1, mild pain; 2, moderate pain; 3,
severe pain; and
4, severe and constant pain). Details of pain measurement after limb
manipu- lation criteria are described in the text and our previous
publications [11-14].

*Significantly

different

compared

to

pretreated

values

(P<0.05).

Figure 3: Effects of placebo or Shilajit (500 mg, bid) on pain after
physical exertion in moderately arthritic dogs. Pain after physical exertion
was graded on a scale of 0-4 (0, no pain; 1, mild pain; 2, moderate pain; 3,
severe pain; and
4, severe and constant pain). Details of pain measurement after physical
exer- tion criteria are described in the text and our previous publications [1114].
Data of physical parameters (body weight, body temperature,
and heart rate are shown in Table 1. Dogs receiving placebo
or Shilajit showed no significant change in any physical
param- eters during the course of this investigation. Table 2
presents serum chemistry data for biomarkers of liver
(bilirubin, ALT, and AST), kidney (BUN and creatinine),
heart and muscle (CK) functions. Dogs receiving placebo or
Shilajit showed no significant change in serum biomarkers
during the study pe- riod of 150 days.

Discussion
Osteoarthritis is prevalent in dogs primarily due to aging, but
it can also be due to obesity, poor nutrition, injury, genetic
pre- disposition and other environmental factors [1, 19-22].
Dogs are also affected by osteoarthritis, a progressive
degenerative disease characterized by loss of cartilage.
Eventually, dogs suf- fer from pain and poor quality of life
due to immobility.
Veterinarians have a wide range of options to treat and
manage osteoarthritis, including surgery, NSAIDS, physical
therapy with acupuncture, nutraceuticals and other disease
modifying agents [13, 14, 23-26, 41].
Currently, glucosamine and chondroitin are the two most
com- monly used nutraceuticals to manage osteoarthritis in
dogs, but their efficacy is limited [24, 35]. In the last ten
years, we have evaluated the therapeutic efficacy and safety of
many nu- traceuticals in arthritic dogs [11-14]. In a most
recent chronic study, we found that Crominex 3+ (a mixture
of trivalent chro- mium, Phyllanthus emblica extract and
purified Shilajit) was very effective in ameliorating arthritic
pain in dogs without causing any untoward events [42].
In the present paper, we report that Shilajit alone at a
higher dose (500 mg, bid) compared to that present in
Crominex 3+ (7.5 mg, bid), is equally effective in reducing
arthritic pain and enhancing the daily activity of dogs
without exerting any side effects. Shilajit administration
ameliorated arthritic pain in all three categories (overall pain,
pain after limb manipulation, and pain after physical
exertion) with maximum effect noted on day 150. Shilajit is
a pale-brown to blackish-brown resin- ous substance and is a
herbo-mineral complex compound that consists of humus
and organic plant (Styrax officinalis, Trifo- lium repens and
others) materials. By having many bioactive minerals and
plant active principles, such as fulvic acid (low molecular
weight substances like uronic acids, phenolic gly- cosides
and amino acids), humic acid (low molecular weight
substances like phenolic acids, anti-oxidants and free radical
scavengers) and free and conjugated dibenzo-alpha-pyrones,
Shilajit is known to exert multiple pharmacological actions.
Some of these actions include antibacterial, anti-arthritic,
anti-ulcerogenic, anti-hyperglycemic, immunomodulatory,
anti-inflammatory, anti-radical, spermatogenic and ovogenic,
anti-aging and energetic properties [28, 38, 40, 43-47]. Shilajit has been used for both preventative health and for treating
many diseases (such as allergies, diabetes, hypertension, loss
of memory, immune dysfunction, arthritis, loss of libido, etc)

for more than 3000 years.
Since Shilajit has multiple bioactive principles, it is highly
likely that it reduced the arthritic pain due to a variety of
bio- chemical and pharmacological mechanisms, including
anti- inflammatory, anti-oxidant, immune-modulatory and
ener- getic properties.

Conclusions
In conclusion, Shilajit is an all-natural supplement which
of- fers significant anti-arthritic properties including
reduction of pain and inflammation. All dogs responded well
to Shilajit ad- ministration without exhibiting any untoward
effects, thereby giving this supplement an edge over many
other nutraceuticals and pharmaceuticals.

Acknowledgements
The authors would like to thank Ms. Robin B. Doss and
Ms. Michelle A. Lasher for their technical assistance in
prepara- tion of this manuscript.

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Anti inflammatory-and-anti-arthritic-efficacy-and-safety-of-purified-shilajit-in-moderately-arthritic-dogs

  • 1. Journal of Veterinary Science & Animal Husbandry Volume 1 | Issue 3 Research Article Open Access Anti-Inflammatory and Anti-Arthritic Efficacy and Safety of Purified Shilajit in Moderately Arthritic Dogs Lawley S1, Gupta RC*1, Goad JT1, Canerdy TD1 and Kalidindi SR2 Murray State University, Breathitt Veterinary Center, Hopkinsville, KY, USA 2 Natreon Inc. New Brunswick, NJ, USA 1 *Corresponding author: Gupta RC, Professor and Head of Toxicology Department, Murray State Univer- sity, Breathitt Veterinary Center, P.O. Box 2000; 715 North Drive, Hopkinsville, KY, 42240-2000, USA; Fax: (270) 886-4295, Tel: (270) 886-3959, E-mail: rgupta@murraystate.edu Citation: Lawley S et al (2013) Anti-Inflammatory and Anti-Arthritic Efficacy and Safety of Purified Shilajit in Moderately Arthritic Dogs. J Vet Sci Anim Husb 1: 302 Received Date: October 31, 2013 Accepted Date: December 12, 2013 Published Date: December 16, 2013 Abstract The objective of this investigation was to evaluate the efficacy and safety of purified Shilajit in moderately arthritic dogs. Ten client- owned dogs in a randomized double-blinded study received either a placebo or Shilajit (500 mg) twice daily for a period of five months. Dogs were evaluated each month for physical condition (body weight, body temperature, heart rate, and respiration rate) and pain associated with arthritis (overall pain, pain from limb manipulation, and pain after physical exertion). Serum samples collected from these dogs were examined each month for biomarkers of liver (bilirubin, ALT, and AST), kidney (BUN and creatinine) heart and mus- cle (creatine kinase) functions. The findings of this study revealed that dogs receiving Shilajit (Group-II) showed a significant (P<0.05) reduction in pain from limb manipulation by day 60, and overall pain and pain after physical exertion by day 120. Maximum pain reduction, using all three criteria, was observed on day 150. Pain level remained significantly unchanged in dogs receiving the placebo. Dogs in either group showed no significant change (P>0.05) in physical parameters or serum markers, suggesting that Shilajit was well tolerated by moderately arthritic dogs. It was concluded that Shilajit significantly (P<0.05) reduced pain in osteoarthritic dogs and markedly improved their daily life without any side effects. Keywords: Purified Shilajit; Osteoarthritis in canine; Shilajit safety; Anti-arthritic nutraceutical Introduction Currently, there are more than 78 million dogs in the US, and every fifth adult dog suffers from arthritis [1]. Dogs suffer more frequently from osteoarthritis (a chronic and progres- sive disease) than any other form of arthritis. Osteoarthritis, also known as degenerative joint disease, is a functional disor- der of the joint, characterized by a change in joint shape sec- ondary to a loss of articular cartilage, osteophyte formation, subchondral sclerosis, bone marrow lesions and synovial pro- liferation, with subsequent alteration of mechanical proper- ties that result in decreased stability, movement and loading [2-9]. In the early stages of osteoarthritis, there is a progressive depletion of the cartilage proteoglycan leading to a net loss of matrix from the cartilage [10]. This leads to a cascade of negative events, including changes in enzymatic cleavage of proteoglycans and an increase in minor collagen types www.annexpublishers.com leading to structural damage and deterioration of the cartilage. The breakdown of cartilage can be increased with certain enzymes including the matrix metalloproteinase enzymes (MMPs). The severity of cartilage lesions in osteoarthritis can be cor- Volume 1 | Issue 3
  • 2. Journal of Veterinary Science & Animal related with the levels of collegenase (MMP-1) present [7Husbandry 9]. The cascading events lead to increased friction and inflamma- tion in the joints. Dogs suffering with osteoarthritis show stiff- ness of joints, crepitus, pain upon manipulation of the joint, periarticular swelling, palpable effusion, restriction in range of motion, lameness and pain and loss of function [7, 11-16]. The combination of history, physical exam, and radiograph can help diagnose osteoarthritis in a canine patient. Radiographs indicate osteophytosis and subchondral bone sclerosis that de- velops over time in osteoarthritic patients. MRI findings can reveal changes consistent with osteoarthritis in the cartilage [17,18]. 2 There are multiple factors which influence dogs to develop ar- thritis, such as genetic predisposition, trauma or injury to the bones or joints, aging, poor nutrition, obesity and environmen- tal factors [1,19]. Often, large breed dogs (e.g. German Shep- herds, Labrador Retrievers, Siberian Huskies, Rottweilers and others) are more prone to develop osteoarthritis than are small breeds [20-22]. www.annexpublishers.com Volume 1 | Issue 3
  • 3. Arthritic dogs show a variety of signs and symptoms, from limping to an inability to walk due to joint stiffness. Pain is the number one complaint [11-14]. The objectives in treating arthritis are to minimize joint pain by reducing the inflamma- tion and slowing the progression of the cartilage damage and to increase joint flexibility. These goals are achieved by employing a variety of pharmaceuticals, nutraceuticals, disease modify- ing agents and physical therapy along with acupuncture [14, 23-26]. Cyclooxygenase (COX) inhibiting non-steroidal antiinflammatory drugs (NSAIDS, such as carprofen, deracoxib, etodolac, firocoxib, ketoprofen, meloxicam, robenacoxib, rofecoxib, tepoxalin, etc) are used with caution as they may cause some adverse effects, such as reduced appetite, vomiting, gastrointestinal bleeding and hepatic and renal damage and dysfunction [23, 27-32]. In a recent prospective (randomized, placebo-controlled, blinded) study, Monteiro-Steagall et al [33] found that the incidence of adverse effects was not statistically different between treated and control dogs. Recently, dog owners and veterinarians have considered man- aging osteoarthritis in dogs with herbs and nutraceuticals. Today, in humans and animals, glucosamine and chondroitin sulfate are the most common nutraceuticals to provide build- ing blocks to repair the cartilage and to minimize and slow down the progression of osteoarthritis to ease inflammatory pain [24,25, 34-37]. Shilajit is a blackish brown exudate from the sedimentary rocks of the pristine himalayan mountains at about 10,000 ft altitude. Purified shilajit is prepared from this exudate by a proprietary extraction process. the main constituents of shila- jit are dibenzo-α-pyrones(DBPs), DBPchromoproteins and fulvic acids with DBP core. On the other hand, alluvial(plant based) fulvic acids do not have DBP core and do not have the same therapeutic efficacy as shjilajit. In fact, alluvial fulvic ac- ids are used as fertilizers. Shilajit exerts many important roles in biological systems: (1) improves bioavailability of minerals and nutrients, (2) provides electrolytes, (3) detoxifies toxic sub- stances, including heavy metals, (4) improves immune system and (5) exhibits antioxidant properties. Shilajit has been used for more than 3000 years for both preventative health and for treating many diseases in humans, including diabetes, arthritis, hypertension, immune dysfunction and loss of memory [3840]. The present study was undertaken to discover the therapeutic efficacy and safety of purified Shilajit in moderately arthritic dogs. The hypothesis of the present investigation was that purified Shilajit will provide aniinflammatory and anti- arthritic effects in moderately arthritic dogs with minimal side effects. In this paper, we present the findings of Shilajit as an ef- fective and safe supplement that exerts anti-inflammatory and anti-arthritic effects in osteoarhtritic dogs. Shilajit appears very promising and offers an alternative to many other previously reported pharmaceuticals and nutraceuticals. Materials and methods Animals
  • 4. 40-65 pounds, were used in this study. These dogs, based on signs of joint stiffness, lameness and radiographic evidence, had pain at the level of moderate arthritis. Our inclusion cri- teria of dogs for this study excluded those having liver, kidney or heart disease, neoplasia, cancer or any other major disease. Institutional Animal Care and Use Committee (IACUC) ap- proval and owner consents were obtained prior to the initia- tion of this study. Experimental design In a randomized double-blinded study, ten client-owned dogs received a placebo (Group-I) or Shilajit (Group-II). The placebo group contained only two dogs because IACUC advised us to keep to the number of dogs to a minimum due to the fact that arthritic dogs suffer from pain for a period of five months. Moreover, findings from our previous studies revealed that dogs receiving placebo showed no improvement in arthritis pain [11-14]. Eight dogs in GroupII received purified Shilajit (500 mg) twice daily (one capsule before morning meal and one capsule before evening meal) for a period of five months. None of the dogs received any treatment or supplement for 3 to 4 weeks prior to the study or during the study period. Pain measurement At pre-determined intervals (i.e. 30 days), each dog was Ten client-owned moderately arthritic dogs, weighing between evalu- ated for overall pain, pain upon limb manipulation and pain af- ter physical exertion, for a period of five months. Overall pain, on a scale of 1-10, was graded as: 0, no pain: 2.5, mild pain: 5, moderate pain: 7.5, severe pain: 10, severe and constant pain. Pain after limb manipulation, on a scale of 0-4, was evaluated as: no pain: 0, mild pain: 1, moderate pain: 2, severe pain: 3, severe and constant pain: 4. Pain after physical exertion, on a scale of 0-4, was evaluated as: no pain: 0, mild pain: 1, moder- ate pain: 2, severe pain: 3, and severe and constant pain: 4. The physical examination of each limb started with the forelimbs and ended with the rear limbs. The evaluation focused on ma- nipulation of the limbs in a forward, backward and circular motion. Three main joints in each limb included for evalua- tion were the shoulder joint, knee joint and stifle joint. Pop- ping and cracking of the joint as well as vocal pain were noted for each canine. Detailed criteria of the measurement of pain are provided in our earlier publications [11-14]. The present investigation was carried out on moderately arthritic dogs. A moderately arthritic dog exhibits overall pain of about 5 on a scale of 1-10; pain upon limb manipulation about 2 on a scale of 1-4; and pain after physical exertion about 2 on a scale of 1-4. Physical examination On a monthly basis, dogs were given a full physical examination for body weight, body temperature and heart rate. Normal values for these parameters are mentioned below Table 1.
  • 5. 3 Journal of Veterinary Science & Animal Husbandry Parameter Unit Placebo/Treated Day 0 Day 30 Day 60 Day 90 Day 120 Day 150 Body Weight lb Placebo 54.1±11.05 54.0±10.8 53.4±9.85 54.6±8.95 53.6±9.95 53.7±9.5 Treated 54.5±3.37 54.9±3.53 54.4±3.26 54.9±3.16 54.5±3.22 54.9±3.27 Heart Rate Beats/min Placebo 135±15 120±0 115±5 90±10 98±2 99±1 Treated 106.8±4.96 117.6±5.89 96.5±6.47 102.4±3.40 102.8±4.60 108.9±2.81 Body Temperature ºF Placebo 100.3±5.41 100.3±0.2 99.9±0.65 100.3±0.2 100.4±0.3 100.1±1.2 Treated 100.1±0.43 99.6±0.26 99.6±0.23 99.6±0.29 100.9±0.30 100.4±0.20 Values are means±SEM; Normal heart rate= 70-160 beats per minute; Normal body temperature= 101-102.5 ºF. Table 1: Effects of placebo or Shilajit (500 mg, bid) on physical parameters in moderately arthritic dogs Parameter Unit Placebo/Treated Day 0 Day 30 Day 60 Day 90 Day 120 Day 150 BUN mg/dl Placebo 19.5±3.5 18±1 22±2 21±1 17±1 17±4 Treated 17.1±2.88 15.6±1.78 18.1±2.30 18.1±2.30 20.5±2.85 20.8±2.64 Placebo 0.92±0.16 0.75±0.02 0.72±0.02 0.76±0.09 0.81±0.02 0.82±0.03 Treated 1.00±0.09 0.79±0.03 0.79±0.04 0.79±0.09 0.92±0.05 0.94±0.04 Placebo 0.5±0.3 0.2±0 0.3±0.1 0.35±0.05 0.2±0 0.3±0.1 Treated 0.21±0.04 0.25±0.04 0.23±0.05 0.39±0.09 0.26±0.04 0.31±0.04 Placebo 45.5±23.5 35.5±8.5 28±0 40±12 27±0 57.5±14.5 Treated 47.9±4.54 59.9±8.41 75.1±20.4 57.6±4.59 60.8±7.36 58.6±4.95 Placebo 127.5±39.5 123±33 84±4 104±15 779.5±629.5 166.4±25.7 Treated 198.4±44.7 349±80.9 224.6±66.2 132.0±28.2 246.0±84.8 166.4±25.7 Placebo 28.5±3.5 22±7 19.5±5.5 23.5±5.5 45.5±26.5 92.5±3.5 Treated 27.1±3.26 31.5±3.13 28.4±3.57 28.5±2.35 30.5±3.59 29.1±3.60 Creatinine mg/dl Total bilirubin mg/dl ALT IU/L AST IU/L CK IU/L Values are means±SEM; Normal reference values: BUN (7-26 mg/dl); Creatinine (0.0-1.35 mg/dl); Total bilirubin (0.1-0.6 mg/dl); ALT (10-120 IU/L); AST (15-65 IU/L); CK (60-450 IU/L). Table 2: Effects of placebo or Shilajit (500 mg, bid) on serum biomarkers of liver (bilirubin, ALT, and AST), kidney (BUN and creatinine), heart and skeletal muscle (CK) functions in moderately arthritic dogs Serum biomarkers assays Blood samples were collected from the cephalic vein using a 3 mL syringe with a 22 gauge needle and were stored in a marble top tube. Samples were then spun to collect serum and transferred to a red top tube for evaluation. Serum samples were collected each month and analyzed for liver (bilirubin, ALT, AST), kidney (BUN and creatinine), heart and muscle (CK) functions, using a Beckman AU 480 serum analyzer. The serum sample assay indicated that neither placebo nor Shila- jit produced adverse effects in vital organs of arthritic dogs. Normal reference values of all serum parameters are provided below Table 2. Statistical analysis The data presented are means ± SEM. Statistical significance of difference was determined by analysis of variance (ANOVA) coupled with Tukey-Kramer test (P<0.05) using the NCSS2000 Statistical Analysis and Graphics Software for Windows®. Results In this double-blinded clinical trial, moderately arthritic dogs receiving placebo (Group-I) or purified Shilajit (500 mg) twice
  • 6. Journal of Veterinary Science & Animal daily for a period of five months, were evaluated for Husbandry physical parameters, pain level and serum biomarkers of liver, kidney, heart and muscle functions. On a monthly basis, each dog was examined for pain level (overall pain, pain after limb manipulation and pain after physical exertion) and the data are shown in Figures 1-3. While evaluating overall pain, the key points were to observe the dog’s gait, range of motion, ability to sit or lie down, ability to rise from a sitting position, ability to rise from a lying position, and posture while standing. Group-II dogs receiving Shilajit showed significant (P<0.05) reduction in overall pain by day 120 (3.00±0.27) compared to day 0 (5.38±0.75). The maximum reduction in overall pain was noted on day 150 (2.38±0.32). Dogs receiving placebo showed no significant change in overall pain. Pain after limb manipulation was measured in each limb of the dog (placed in lateral recumbency) for flexibility, joint integrity, crepitance and vocalization. The pain level was sig- nificantly reduced by day 60 (1.63±0.18) and the trend contin- ued until day 150 when the pain level was maximally reduced (1.25±0.31) (Figure 2). Pain after limb manipulation remained significantly unchanged in Group-1 (placebo) dogs. Figure 3 shows the data of pain after physical exertion in dogs receiving placebo or Shilajit. The canines were evaluated for pain after three minutes of jogging. After jogging, pain level was assessed 4
  • 7. based on the dog’s unwillingness to move, body position, limp- ing, flexibilty, joint integrity, crepitance and vocalization. Sig- nificant reduction in pain after physical exertion was noted on day 120 (1.37±0.26) compared to day 0 (2.63±0.26) in dogs re- ceving Shilajit. The maximum reduction in pain was observed on day 150 (1.25±0.28). Dogs on placebo showed no signifi- cant change in pain level. *Significantly (P<0.05). different compared to pre-treated values Figure 1: Effects of placebo or Shilajit (500 mg, bid) on overall pain in mod- erately arthritic dogs. Overall pain was graded on a scale of 0-10 (0, no pain; 2.5, mild pain; 5, moderate pain; 7.5, severe pain; and 10, severe and constant pain). Details of overall pain measurement criteria are described in the text and our previous publications [11-14]. *Significantly (P<0.05). different compared to pretreated values Figure 2: Effects of placebo or Shilajit (500 mg, bid) on pain from limb manip- ulation in moderately arthritic dogs. Pain from limb manipulation was graded on a scale of 0-4 (0, no pain; 1, mild pain; 2, moderate pain; 3, severe pain; and 4, severe and constant pain). Details of pain measurement after limb manipu- lation criteria are described in the text and our previous publications [11-14]. *Significantly different compared to pretreated values (P<0.05). Figure 3: Effects of placebo or Shilajit (500 mg, bid) on pain after physical exertion in moderately arthritic dogs. Pain after physical exertion was graded on a scale of 0-4 (0, no pain; 1, mild pain; 2, moderate pain; 3, severe pain; and 4, severe and constant pain). Details of pain measurement after physical exer- tion criteria are described in the text and our previous publications [1114].
  • 8. Data of physical parameters (body weight, body temperature, and heart rate are shown in Table 1. Dogs receiving placebo or Shilajit showed no significant change in any physical param- eters during the course of this investigation. Table 2 presents serum chemistry data for biomarkers of liver (bilirubin, ALT, and AST), kidney (BUN and creatinine), heart and muscle (CK) functions. Dogs receiving placebo or Shilajit showed no significant change in serum biomarkers during the study pe- riod of 150 days. Discussion Osteoarthritis is prevalent in dogs primarily due to aging, but it can also be due to obesity, poor nutrition, injury, genetic pre- disposition and other environmental factors [1, 19-22]. Dogs are also affected by osteoarthritis, a progressive degenerative disease characterized by loss of cartilage. Eventually, dogs suf- fer from pain and poor quality of life due to immobility. Veterinarians have a wide range of options to treat and manage osteoarthritis, including surgery, NSAIDS, physical therapy with acupuncture, nutraceuticals and other disease modifying agents [13, 14, 23-26, 41]. Currently, glucosamine and chondroitin are the two most com- monly used nutraceuticals to manage osteoarthritis in dogs, but their efficacy is limited [24, 35]. In the last ten years, we have evaluated the therapeutic efficacy and safety of many nu- traceuticals in arthritic dogs [11-14]. In a most recent chronic study, we found that Crominex 3+ (a mixture of trivalent chro- mium, Phyllanthus emblica extract and purified Shilajit) was very effective in ameliorating arthritic pain in dogs without causing any untoward events [42]. In the present paper, we report that Shilajit alone at a higher dose (500 mg, bid) compared to that present in Crominex 3+ (7.5 mg, bid), is equally effective in reducing arthritic pain and enhancing the daily activity of dogs without exerting any side effects. Shilajit administration ameliorated arthritic pain in all three categories (overall pain, pain after limb manipulation, and pain after physical exertion) with maximum effect noted on day 150. Shilajit is a pale-brown to blackish-brown resin- ous substance and is a herbo-mineral complex compound that consists of humus and organic plant (Styrax officinalis, Trifo- lium repens and others) materials. By having many bioactive minerals and plant active principles, such as fulvic acid (low molecular weight substances like uronic acids, phenolic gly- cosides and amino acids), humic acid (low molecular weight substances like phenolic acids, anti-oxidants and free radical scavengers) and free and conjugated dibenzo-alpha-pyrones, Shilajit is known to exert multiple pharmacological actions. Some of these actions include antibacterial, anti-arthritic, anti-ulcerogenic, anti-hyperglycemic, immunomodulatory, anti-inflammatory, anti-radical, spermatogenic and ovogenic, anti-aging and energetic properties [28, 38, 40, 43-47]. Shilajit has been used for both preventative health and for treating many diseases (such as allergies, diabetes, hypertension, loss of memory, immune dysfunction, arthritis, loss of libido, etc) for more than 3000 years.
  • 9. Since Shilajit has multiple bioactive principles, it is highly likely that it reduced the arthritic pain due to a variety of bio- chemical and pharmacological mechanisms, including anti- inflammatory, anti-oxidant, immune-modulatory and ener- getic properties. Conclusions In conclusion, Shilajit is an all-natural supplement which of- fers significant anti-arthritic properties including reduction of pain and inflammation. All dogs responded well to Shilajit ad- ministration without exhibiting any untoward effects, thereby giving this supplement an edge over many other nutraceuticals and pharmaceuticals. Acknowledgements The authors would like to thank Ms. Robin B. Doss and Ms. Michelle A. Lasher for their technical assistance in prepara- tion of this manuscript. References 1) Arthritis Foundation (2013). Spotting Arthritis in Spot. Retrieved 09 2013, 02, from Arthritis Foundation. 2) Vaughan-Scott T, Taylor JH (1997). The pathophysiology and medical man- agement of canine osteoarthritis. J S Afr Vet Assoc. 68: 21-25. 3) Bellamy N, Carr A, Dougados M, Shea B, Wells G (2001). Towards a defini- tion of “differences” in osteoarthritis. J Rheumatol. 28: 427-430. 4) Lennon E, Marcellin-Little D (2005). Canine Osteoarthritis. Retrieved Au- gust 5, 2013 from Arthritis M.D. 5) Burns K (2006). Research targets and conditions of older cats and dogs. J Am Vet Med Assoc. 229: 482-483. 6) Centers for Disease Control and Prevention (CDC) Arthritis-BasicDefini- tion-Osteoarthritis. Retrieved June 18, 2010. 7) Renberg WC (2005). Pathophysiology and management of arthritis. Vet Clin Small Anim. 35: 10731091. 8) Dore D, Martens A, Quinn S, Ding C, Winzenberg C, et al.(2010) Bone marrow lesions predict site-specific cartilage defect development and volume loss: a prospecitve study in older adults. Arthr Res Ther. 12: R222. 9) Umlauf D, Frank S, Pap T, Bertrand J (2010) Cartilage, biolog y, pathology, and repair. Cell Mol Life Sci. 67: 4197-4211. 10) Reid DM, Miller GC (2008). Clinical Trails in Rheumatoid Arthritis and Osteoarthritis.Aberdeen, UK: Springer-Verlag London Limited. 11) Deparle LA, Gupta RC, Canerdy TD, Goad JT, D’Altilio M, et al. (2005). Efficacy and safety of glycosylated undenatured type-II collagen (UC-II) in therapy of arthritic dogs. J Vet Pharmacol Ther. 28: 385-390. 12) D’Altilio M, Peal A, Alvey M, Simms C, Curtsinger A, et al. (2007). Thera- peutic efficacy and safety of undenatured type II collagen singly or in combi- nation with glucosamine and chondroitin in arthritic dogs. Toxicol Mechan Methods.17:189-196. 13) Peal A, D’Altilio M, Simms C, Alvey M, Gupta RC, et al. (2007). Thera- peutic efficacy and safety of undenatured type-II collagen (UC-II) alone or in combination with (-) hydroxycitric acid and chromemate in arthritic dogs. J Vet Pharmacol Ther. 30: 275-278. 14) Gupta RC, Canerdy TD, Lindley J, Konemman M, Minniear J, et al. (2012) Comparative therapeutic efficacy and safety of type-II collagen (UC- II), glu- cosamine and chondroitin in arthritic dogs: pain evaluation by ground force plate. J Anim Physiol Anim Nutri. 96: 770-777. 15) Osteoarthritis in Dogs- Signs and Symptoms of Arthritis and Osteoarthritis. (2012) Retrieved August 8, 2013, from Osteoarthritis in Dogs.
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  • 11. 44) Bhattacharya SK, Sen AP, Ghosal S (1995). Effects of Shilajit on biogenic free radicals. Phytother Res 9: 56-59. 45) Tripathi YB, Shukla S, Chaurasia S, Chaturvedi S (1996). Antilipid peroxi- dative property of Shilajit. Phytother Res. 10: 269-270. 46) Park JS, Kim GY, Han K (2006). The spermatogenic and ovogenic effects of chronically administered Shilajit to rats. J Ethnopharmacol. 107: 349-353. 47) Meena H, Pandey HK, Arya MC, Ahmed Z (2010). Shilajit: A panacea for high-altitude problems. Int J Ayurveda Res, 1 (1): 37-40.